BRPI0807718B1 - COMPOSITION INCLUDING STABLE DIGESTIVE ENZYME, USE OF THE SAME, DOSAGE FORM, PACKAGING AND PREPARATION PROCESS OF A PANCRELIPASE COMPOSITION - Google Patents

Abstract

composition comprising stable digestive enzyme, use thereof, dosage form, packaging and process for preparing a pancrelipase composition. The present invention relates to compositions of the present invention, comprising at least one digestive enzyme (e.g. pancrelipase), are useful for treating or preventing disorders associated with digestive enzyme deficiencies. the compositions of the present invention may comprise a plurality of coated particles, each of which is comprised of a core coated with an enteric coating comprising at least one enteric polymer and 4-10% of at least one alkalizing agent or has moisture contents of about 3% or less, water activities of about 0.6 or less, or exhibit an activity loss of no more than about 15% after six months of accelerated stability testing.

Description

Cross Reference to Related Orders

The present application claims priority to US Provisional Patent Application No. 60/902,091 filed February 20, 2007, US Provisional Patent Application No. 60/902,093 filed February 20, 2007 and US Provisional Patent Application No. 60/902,092 filed February 20, 2007, the descriptions of which are incorporated herein by reference in their entirety for all purposes.

Background of the Invention

In cases of pancreatic insufficiency, pancrelipase and other pancreatic enzyme products (PEPs) can be administered to at least remedy the enzyme deficiency caused by various diseases that affect the pancreas, such as pancreatitis, pancreatectomy, cystic fibrosis, etc. The use of pancreatic enzymes in the treatment of pancreatic insufficiency is an essential part of the therapy of patients afflicted with cystic fibrosis. Without these supplements, patients are severely nutritionally impaired. This nutritional impairment can be life threatening if left untreated, particularly in the case of babies.

The drug substance pancrelipase is primarily a combination of three enzyme classes: lipase, protease and amylase, along with their various cofactors and coenzymes. These enzymes are naturally produced in the pancreas and are important in the digestion of fats, proteins and carbohydrates. Pancrelipase is typically prepared from porcine pancreatic glands, although other sources may also be used, for example those described in US 6,051,220, US 2004/0057944, 2001/0046493 and WO 2006044529, each incorporated herein by way of reference. Enzymes catalyze the hydrolysis of fats into glycerol and fatty acids, starch into dextrin and sugars, and protein into amino acids and derived substances.

Pancreatic enzymes show optimal activity under almost neutral and slightly alkaline conditions. Under gastric conditions, pancreatic enzymes can be inactivated with a resulting loss in biological activity. Thus, exogenously administered enzymes are generally protected against gastric inactivation and remain intact during their transit through the stomach and into the duodenum. While it is desirable to coat pancreatic enzymes, uncoated preparations are also commercially available.

Pancreatic lipases are the most sensitive to gastric inactivation and are the single most important enzymes in the treatment of malabsorption. Lipase activity is typically monitored to determine the stability of a lipase-containing enzyme composition.

After passing through the stomach, enzymes must be released into the duodenum within 5-30 minutes, as digestion by pancreatic enzymes and absorption of metabolites takes place mainly in the upper segment of the intestine, although digestion and absorption can take place through the Gl transit. Pancreatic cells have typically been coated with an enteric coating polymer, which protects the enzyme composition against the acidic environment of the stomach and then provides release of the enzyme in the intestine.

Conventional pancreatic enzyme preparations are inherently unstable and do not have the half-life associated with pharmaceuticals approved for oral use. The activity of pancreatic enzyme preparations is typically determined based on lipase activity, which is one of the most sensitive enzymes for loss of enzyme activity during storage. Commercially available digestive enzyme compositions show a loss of lipase activity over time of up to about 35% or more. In order to compensate for losses in enzyme activity during storage and to ensure that the product delivers the potency claimed by the label at the end of its shelf life, manufacturers typically overfill dosage forms from 5% to 60% and Current USP specifications for Pancrelipase Delayed Release Capsules allow Pancrelipase equivalent to no less than 90% and no more than 165% of labeled lipase activity.

In practice this means that patients and prescribers are unable to judge the dosage concentration accurately, with the practical result that the appropriate dosage needs to be determined empirically for each new prescription. Patients with disorders of exocrine pancreatic insufficiency rely on these drugs to provide the enzymes they need to properly digest food. If the label contains an inaccurate statement about the potency of the particular product, then the patient is at risk of receiving too much or too little of the drug.

Thus, it would be desirable to provide a stable digestive enzyme composition capable of maintaining the activity necessary for the expected shelf life of the enzyme preparation, without relying on overfilling the dosage form.

Invention Summary

The present invention relates to stable digestive enzyme compositions and dosage forms and methods for producing stable enzyme compositions and dosage forms. More particularly, the present invention relates to coated enzyme compositions and enteral dosage forms that exhibit minimal loss of activity under typical storage conditions.

In one embodiment, the present invention provides a composition comprising at least one digestive enzyme, where the composition has a moisture content of about 3% or less.

In another embodiment, a composition of the present invention comprises at least one digestive enzyme, where the composition has a water activity of about 0.6 or less.

In another embodiment, a composition of the present invention comprises at least one stabilized digestive enzyme, where the at least one stabilized digestive enzyme exhibits a loss of activity of no more than about 15% after six months of accelerated stability testing.

In yet another embodiment, the present invention provides a dosage form such as a tablet or capsule filled with the composition of the present invention.

In yet another embodiment, a composition of the present invention further comprises the at least one coating coated digestive enzyme, wherein the coating comprises an enteric polymer and optionally at least one inorganic material.

In yet another embodiment, the present invention provides a package comprising a sealed container made of moisture resistant material, a desiccant and at least one dosage form of the present invention, wherein the desiccant and at least one dosage form are within the container. tight.

In yet another embodiment, the present invention provides a method of treating or preventing a disorder associated with digestive enzyme deficiency comprising administering a composition of the present invention to a mammal in need thereof.

In yet another embodiment, the present invention provides a method of preparing a composition of the present invention. In one embodiment, the method comprises coating particles of the at least one digestive enzyme in an atmosphere having a moisture content of about 3.6 g of water per m3 or less, with a coating comprising an enteric polymer and at least one material. inorganic, thereby forming a plurality of delayed release particles.

Detailed Description of the Invention

One aspect of the present invention pertains to a stabilized digestive enzyme composition. The term "stabilized digestive enzyme" means a digestive enzyme that maintains substantial enzyme activity after long-term storage. The term "digestive enzyme" means an enzyme in the alimentary tract that breaks down food components so that they can be absorbed by the body.

Non-limiting classes of digestive enzymes suitable for use in the present invention include lipases, amylases and proteases. Non-limiting examples of digestive enzymes include pancrelipase (also referred to as pancreatin), lipase, colipase, trypsin, chymotrypsin, chymotrypsin B, pancreatopeptidase, carboxypeptidase A, carboxypeptidase B, glycerol ester hydrolase, phospholipase, sterol ester hydrolase, ribokinase, elastase , deoxyribonuclease, α-amylase, papain, chymopapain, glutenase, bromelain, ficin, β-amylase, cellulase, β-galactosidase, lactase, sucrase, isomaltase and mixtures thereof.

In one embodiment of the present invention, the stabilized digestive enzyme is a pancreatic enzyme. The term "pancreatic enzyme" as used herein refers to any of the types of enzyme present in pancreatic secretion, such as amylase, lipase, protease, or mixtures thereof, or any extractive of pancreatic origin having enzymatic activity, such as pancreatin . The pancreatic enzyme can be obtained through extraction from the pancreas, produced artificially or obtained from sources other than the pancreas, such as from microbes, plants or other animal tissues.

In another embodiment of the present invention, the stabilized digestive enzyme is pancrelipase. The term "pancrelipase" or "pancreatin" means a mixture of several types of enzymes, including amylase, lipase and protease enzymes. Pancrelipase is commercially available, for example, from Nordmark Arzneimittel GmbH or Scientific Protein Laboratories LLC.

In one embodiment of the compositions of the present invention, the stabilized digestive enzyme comprises a lipase. The term "lipase" refers to an enzyme that catalyzes the hydrolysis of lipids to glycerol and simple fatty acids.

Examples of suitable lipases for the present invention include, but are not limited to, animal lipase (for example pig lipase), bacterial lipase (for example Pseudomonas lipase and/or Burkholderia lipase), fungal lipase, lipase from plant, recombinant lipase (e.g., produced through recombinant DNA technology by a suitable host cell, selected from any of bacterial, yeast, fungal, plant, insect, or mammalian host cells in culture, or recombinant lipases that include a amino acid sequence that is homologous or substantially identical to a naturally occurring sequence, lipases encoded by a nucleic acid that is homologous or substantially identical to a naturally occurring lipase encoding nucleic acid, etc.), chemically modified lipase or mixtures thereof .

In another embodiment of the compositions of the present invention, the stabilized digestive enzyme comprises an amylase. The term "amylase" refers to glycoside hydrolase enzymes that break down starch, for example α-amylases, β-amylases, y-amylases, acidic α-glycosidases, salivary amylases such as ptyalin etc

Suitable amylases for use in the compositions of the present invention include, but are not limited to, animal amylases, bacterial amylases, fungal amylases (e.g., Aspergillus amylase and preferably is Aspergillus oryzae amylase), plant amylases, amylases recombinants (for example, produced through recombinant DNA technology by a suitable host cell, selected from any of bacterial, yeast, fungi, plant, insect or mammalian host cells, or recombinant amylases that include an amino acid sequence that is homologous or substantially identical to a naturally occurring sequence, amylases encoded by a nucleic acid which is homologous or substantially identical to a naturally occurring amylase encoding nucleic acid, etc.), chemically modified amylases or mixtures thereof.

In another embodiment of the compositions of the present invention, the stabilized digestive enzyme comprises a protease. The term "protease" generally refers to enzymes (for example, proteinases, peptidases or proteolytic enzymes) that break peptide bonds between amino acids in proteins. Proteases are generally identified by their catalytic type, for example, aspartic acid peptidases, cysteine (thiol) peptidases, metallopeptidases, serine peptidases, threonine peptidases, alkaline or semi-alkaline, neutral proteases and peptidases of unknown catalytic mechanism.

Non-limiting examples of proteases suitable for use in the compositions or oral dosage forms of the present invention include serine proteases, threonine proteases, cysteine proteases, aspartic acid proteases (e.g., plasmepsin), metalloproteases, glutamic acid proteases, etc., in In addition, proteases suitable for use in the compositions or oral dosage forms of the present invention include, but are not limited to animal proteases, bacterial proteases, fungal proteases (e.g., an Aspergillus melleus protease), plant proteases, recombinant proteases (for example, for example, produced through recombinant DNA technology by a suitable host cell, selected from any of the host cells of bacteria, yeast, fungi, plant, insect or mammal in culture, or recombinant proteases that include an amino acid sequence that is homologous or substantially identical to a naturally occurring sequence, acid-encoded proteases. of the nucleic acid which is homologous or substantially identical to a nucleic acid encoding naturally occurring protease, etc.), chemically modified proteases or mixtures thereof.

The compositions or oral dosage forms of the present invention can comprise one or more lipases (i.e., a lipase, or two or more lipases), one or more amylases (e.g., an amylase, or two or more amylases), one or more more proteases (i.e. one protease, or two or more proteases), mixtures of one or more lipases with one or more amylases, mixtures of one or more lipases with one or more proteases, mixtures of one or more amylases with one or more proteases or mixtures of one or more lipases with one or more amylases and one or more proteases.

In one embodiment, the digestive enzyme is a porcine pancreatic extract comprising various lipases (eg, lipase, colipase, phospholipase A2, cholesterol esterase), proteases (eg, trypsin, chymotrypsin, carboxypeptidases A and B, elastase , kininogenase, trypsin inhibitor), amylases and optionally nucleases (ribonuclease, deoxyribonuclease). In another embodiment, the digestive enzyme is substantially similar to human pancreatic fluid. In yet another embodiment, the digestive enzyme is pancrelipase USP. In yet another embodiment, the digestive enzyme is pancrelipase ESP having a lipase activity of 69-120 U USP/mg, amylase activity greater than or equal to 216 USP/mg, protease activity greater than or equal to 264 U USP/mg and total protease activity greater than or equal to 264 U USP/mg.

Lipase activities in the compositions or oral dosage forms of the present invention can be about 4500-25,000 IU, for example, about 4500-5500 IU, about 9000-11,000 IU, about 13,500-16,500 IU and about 18,000- 22,000 IU. Amylase activities in the compositions or oral dosage forms of the present invention can be about 8100-180,000 IU, for example, about 8000-45,000 IU, about 17,000-90,000 IU, about 26,000-135,000 IU, about 35,000- 180,000 IU. Protease activities in the compositions or oral dosage forms of the present invention can be about 8,000-134,000 IU, for example, about 8000-34,000 IU, 17,000-67,000 IU, 26,000-100,000 IU, 35,000-134,000 IU. In one embodiment, lipase activity ranges from about 4500-5500 IU, amylase activity ranges from about 8000-45,000 IU, and protease activity ranges from about 8000-34,000 IU UI. In another embodiment, lipase activity ranges from about 9,000-11,000 IU, amylase activity ranges from about 17,000-90,000 IU, and protease activity ranges from about 17,000-67,000 IU. UI. In yet another modality, lipase activity ranges from about 13,500-16,500 IU, amylase activity ranges from about 26,000-135,000 IU, and protease activity ranges from about 26,000- 100,000 IU. In yet another modality, lipase activity ranges from about 18,000-22,000 IU, amylase activity ranges from about 35,000-180,000 IU, and protease activity ranges from about 35,000- 134,000 IU.

The lipase:protease:amylase ratios in the compositions or oral dosage forms of the present invention can range from about 1:10:10 to about 10:1:1 or about 1.0:1.0:0 .15 (based on enzyme activities). The amylase/lipase ratio in the compositions or oral dosage forms of the present invention can range from about 1.8-8.2, e.g., about 1.9-8.2, and about 2. 0-8.2. The proteinase/lipase ratio in the compositions in the oral dosage forms of the present invention can range from about 1.8-6.2, e.g., about 1.9-6.1, and about 2.0-6.1.

sições ou formas de dosagem orais da presente invenção pode ser cerca de 20-100%, 20-90%, 20-80%, 20-70%, 20-60%, 20-50%, 20-40%, 20-30% ou cerca de 20%, cerca de 30%, cerca de 40%, cerca de 50%, cerca de 60%, cerca de 70%, cerca de 80%, cerca de 90% ou cerca de 100%. Em uma modalidade, a quantidade total de enzimas digestivas é 60-90%. Em outra modalidade, a quantidade total de enzimas digestivas (por exemplo, pancrelipase) é cerca de 68-72%.In another embodiment, the lipase, protease and amylase activities can be those described in Table A below: Table A

Oral dosage forms or compositions of the present invention may be about 20-100%, 20-90%, 20-80%, 20-70%, 20-60%, 20-50%, 20-40%, 20- 30% or about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100%. In one modality, the total amount of digestive enzymes is 60-90%. In another embodiment, the total amount of digestive enzymes (eg pancrelipase) is about 68-72%.

In one embodiment, the compositions or oral dosage forms of the present invention, comprising at least one digestive enzyme, have a moisture content of about 3% or less, about 2.5% or less, about 2% or less , about 1.5% or less, or about 1% or less, inclusive of all subband ranges in between (ie, any of about 2.5% to 3%, 2% to 3%, 1 .5% to 3%, 1% to 3%, 2% to 2.5%, 1.5% to 2.5%, 1% to 2.5%, 1.5% to 2%, 1% to 2%, 1% to 1.5%, etc). Oral compositions or dosage forms of the present invention, maintained at a low moisture content, have been found to be substantially more stable compared to conventional compositions maintained at higher moisture contents, e.g., about 3% or more.

The term "moisture content", also referred to as "water content", means the amount of water that a composition contains. For compositions that do not change volume with changing moisture content, moisture content can be expressed volumetrically (ie, by volume) as the ratio of the mass of moisture to the dry volume of the material. For volume changing compositions with changing moisture content, the moisture content may be expressed gravimetrically (i.e., by weight) as the mass of water removed upon drying per unit dry mass of the specimen. Determination of moisture content can be accomplished by any conventional methods known in the field. For example, moisture content can be determined through chemical titration, such as Karl Fischer titration, where a sample is dissolved in an electrochemical titration cell. Sample water is consumed in an electrochemical reaction whose end point is measured potentiometrically, thus providing a direct measure of the amount of water in the sample. Alternatively, relatively simple thermogravimetric methods can be used such as "Loss on Drying" (LoD), where the mass of a sample is measured before and after controlled drying. The loss of mass after drying is attributed to the loss of moisture. Commercially available moisture analyzers (eg available from Mettler Toledo, Sartorius AG, etc.) can also be used to determine moisture content. The moisture content of the compositions or oral dosage forms of the present invention can be measured by any suitable method known in the art, for example, LoD.

In another embodiment, the compositions or oral dosage forms of the present invention, comprising at least one digestive enzyme, have a water activity of about 0.6 or less, about 0.5 or less, about 0.4 or less, about 0.3 or less, about 0.2 or less, or about 0.1 or less, inclusive of all ranges and subranges in between (ie, any one from about 0.5 to 0.6, 0.4 to 0.6, 0.3 to 0.6.0.2 to 0.6, 0.1 to 0.6, 0.4 to 0.5, 0.3 to 0. 5, 0.2 to 0.5, 0.1 to 0.5, 0.3 to 0.4, 0.2 to 0.4, 0.1 to 0.4, 0.2 to 0.3, 0.1 to 0.3, 0.1 to 0.2, etc). Compositions or oral dosage forms of the present invention, maintained at a lower water activity, were found to be substantially more stable compared to conventional digestive enzyme compositions maintained at higher water activity levels.

Water activity, also referred to as "aw", is the relative availability of water in a substance. As used herein, the term "water activity" is defined as the vapor pressure of water in a sample divided by the vapor pressure of pure water at the same temperature. Pure distilled water has a water activity of exactly one. Water activity is temperature dependent. That is, water activity changes as temperature changes. In the present invention, water activity is measured at a temperature ranging from about 0°C to about 50°C, preferably from about 10°C to about 40°C.

The water activity of a product can be determined by measuring the relative humidity of the air surrounding the equilibrium sample. Thus, measurement of water activity in a sample is typically performed in an enclosed (usually isolated) space, where this equilibrium can take place. At equilibrium, the water activity of the sample and the relative humidity of the air are equal, and so a measurement of the equilibrium relative humidity (ERH) of the air in the chamber provides a measure of the water activity of the sample. At least two different types of water activity instruments are commercially available. One type of water activity instruments use chilled mirror dew point technology (eg, AquaLab® water activity meters available from Decagon Devices, Inc.) while others measure relative humidity with sensors that change electrical resistance or capacitance (eg water activity meters available from Rotronic). The water activity of the compositions or oral dosage forms of the present invention can be measured by any suitable method known in the art.

In another embodiment, the compositions or oral dosage forms of the present invention, comprising at least one stabilized digestive enzyme, exhibit a loss of enzyme activity of no more than about 25%, no more than about 20%, no more than about 15%, no more than about 12%, no more than about 10%, no more than about 8% or no more than about 5%, after six months of testing. accelerated stability.

The term "accelerated stability test" or "accelerated storage test" refers to test methods used to simulate the effects of relatively long-term storage conditions on enzyme activity, which can be performed in a relatively short time. Accelerated stability testing methods are known in the art to be a reliable alternative to real-time stability testing, and can accurately predict the shelf life of biological products. Such "accelerated stability testing" conditions are known in the art and are in accordance with the International Conference for Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use: Stability Testing of New Drug Substances and Products Q1A, incorporated herein by reference in its entirety.

One accelerated stability test method comprises storing digestive enzyme composition samples in a sealed Nialene bag (nylon, aluminum, polyethylene laminate; GOGLIO SpA, Milan) at 40°C/75% relative humidity for 6 months.

After storage (or periodically during storage) the enzyme activity of samples can be tested using conventional methods for assaying digestive enzyme activity (eg, United States Pharmacopoeia, Pancrelipase: Assay for lipase activity; incorporated herein by reference in its totality).

The compositions or oral dosage forms of the present invention may also further comprise one or more stabilizers which increase or improve the stability of the compositions or oral dosage forms of the present invention. Non-limiting examples of suitable stabilizers include proline, trehalose, dextran, maltose, sucrose, mannitol, polyols, silica gel, aminoguanidine, pyridoxamine, anhydrous metal salts such as magnesium oxide, sodium hydrogen carbonate, calcium oxide , aluminum oxide and mixtures thereof. The one or more stabilizers may have a moisture content of about 3% or less and/or a water activity of 0.6 or less.

Non-limiting examples of suitable forms of trehalose that can be used in the compositions or oral dosage forms of the present invention include trehalose dihydrate (TD), amorphous trehalose (AT), anhydrous trehalose (e.g., anhydrous amorphous trehalose (AAT). ), anhydrous crystalline trehalose (ACT)). Anhydrous trehalose powder may contain any AAT and/or ACT. As used herein, the term "trehalose" refers to any physical form of trehalose, including anhydrous, partially hydrated, fully hydrated, and mixtures and solutions thereof. The term "anhydrous trehalose" refers to any physical form of trehalose containing less than 2% water. Anhydrous forms of trehalose can contain from 0-2% water. Amorphous trehalose contains about 2-9% water and trehalose dihydrate contains about 9-10% water. Anhydrous trehalose can be repaired as described in PCT/GB97/00367, incorporated herein by reference in its entirety. In one embodiment, the compositions or oral dosage forms of the present invention comprise one or more stabilized digestive enzymes and anhydrous trehalose.

The amount of anhydrous trehalose (AAT or ACT) in the composition of the present invention can be in the range of about 5-50%, 5-40%, 5-30%, 5-20%, 5-15%, 5-10 %, 7-15% or about 5%, about 7%, about 10%, about 15% or about 20%.

Anhydrous trehalose can be incorporated into the compositions or oral dosage forms of the present invention as a powder. The particle size of anhydrous trehalose powder can be in the range of about 2-2000 µm.

Compositions or oral dosage forms of the present invention comprising one or more stabilized digestive enzymes and anhydrous trehalose impart improved enzyme stability. Anhydrous tre-alose is believed to stabilize the compositions or oral dosage forms of the present invention by absorbing or sequestering moisture from ambient moisture, or residual moisture from manufacturing or within the formulation itself.

Depending on the intended use and requirement of the compositions, the weight ratio of stabilized digestive enzyme to stabilizer ranges from about 99:1 to 80:20. The stabilizer can be incorporated into oral compositions or dosage forms of the present invention by dry or wet mixing of at least one stabilized digestive enzyme with at least one stabilizer. In one embodiment, one or more stabilized digestive enzymes is dry blended with one or more stabilizers. In another embodiment, one or more stabilized digestive enzymes is wet mixed with one or more stabilizers.

In addition to the stabilized digestive enzyme and/or stabilizer(s), the compositions or oral dosage forms of the present invention can further comprise one or more pharmaceutically acceptable excipients. The term "excipients" includes other pharmaceutically acceptable ingredients added to the active component(s) of a composition (e.g., stabilized digestive enzymes) in order to improve processing, stability, palatability, etc. Non-limiting examples of suitable excipients include binders, stabilizers, disintegrants, lubricants, non-sticks, pharmaceutically acceptable diluents and mixtures thereof, etc. It will be understood by those skilled in the art of pharmaceutical formulations that a particular excipient can perform multiple functions in the composition. So, for example, a binder can also function as a diluent, etc. Excipients can have a moisture content of about 3% or less and/or water activity of about 0.6 or less.

Non-limiting examples of suitable binders include starches, sugars (eg lactose), sugar alcohols (eg xylitol, sorbitol, matitol), cellulose (eg microcrystalline cellulose), modified celluloses ( for example, hydroxypropylcellulose, sodium carboxymethylcellulose), alginic acid, polyvinyl pyrrolidone (povidone) and mixtures thereof. Non-limiting examples of suitable disintegrants include dibasic dicalcium phosphate, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, alginic acid, hydroxypropylcellulose, calcium carboxymethylcellulose, sodium carboxymethylcellulose, cross-linked sodium carboxymethylcellulose, intumescent exchange resins ion, alginates, formaldehyde-casein, cellulose, croscarmellose sodium, crospovidone (eg, cross-linked polyvinyl pyrrolidone), microcrystalline cellulose, sodium carboxymethyl starch, sodium starch glycolate, starches (corn starch, rice starch ) and mixtures thereof. Non-limiting examples of suitable lubricants include calcium stearate, magnesium stearate, sodium stearyl fumarate, stearic acid, zinc stearate, talc, waxes, Sterotex®, Stearowet® and mixtures thereof. Non-limiting examples of suitable non-sticks include colloidal silica dioxide, talc and mixtures thereof. Non-limiting examples of suitable diluents include mannitol, sucrose, anhydrous dibasic calcium phosphate, anhydrous dibasic calcium phosphate dihydrate, tribasic calcium phosphate, cellulose, lactose, magnesium carbonate, microcrystalline cellulose and mixtures thereof. Non-limiting examples of suitable stabilizers include trehalose, proline, dextran, maltose, sucrose, mannitol, polyol, silica gel, aminoguanidine, pyridoxamine and mixtures thereof.

In one embodiment, the disintegrant is crospovidone (for example, POLYPLASDONA XL, POLYPLASDONA XL-10). In another embodiment, the disintegrant is croscarmellose sodium (eg, AC-DI-SOL). In another embodiment, the disintegrant is sodium starch glycolate (eg EXPLOTAB, EXPLOTAB CV). In another embodiment, the compositions or oral dosage forms of the present invention can comprise a combination of disintegrants such as microcrystalline cellulose and sodium starch glycolate or croscarmellose sodium and crospovidone.

The amount of disintegrants can range anywhere from about 0.1-30%, 1-30%, 1%-25%, 1%-20%, 1%-15%, 1%-10%, 1%-5%, 5%-10%, 5%-15%, 5%-20%, 5%-25% or 5%-30%. In one embodiment, the amount of disintegrant is about 2%-4% or about 2%-3% or about 2.5%.

Non-limiting examples of suitable diluents include microcrystalline cellulose, starch, calcium phosphate, lactose, sucrose, mannitol, sorbitol and combinations thereof. In one embodiment, the diluent is microcrystalline cellulose (eg, Avicel). In another modality, the thinner is starch. In another embodiment, the diluent is lactose (eg, Pharmatol). In another embodiment, the compositions or oral dosage forms of the present invention can comprise a combination of diluents such as microcrystalline cellulose, starch and lactose.

The amount of diluent can be in the range of about any of 0.1-99%, 1%-30%, 1%-25%, 1%-20%, 1%-15%, 1%-10% , 1%-5%, 5%-10%, 5%-15%, 5%-20%, 5%-25% or 5%-30%. In one embodiment, the amount of thinner is about 2%-5%, 3%-5%, or about 4%.

One or more of the excipients in the compositions or oral dosage forms of the present invention can function as a desiccant to further stabilize the composition. Suitable excipients useful as desiccants include any pharmaceutically acceptable excipient which strongly binds water or reduces the water activity of a composition. For example, the composition of the present invention can include about 1-4% silica gel or about 2.5% silica gel.

Compositions of the present invention may be prepared in any suitable oral dosage form. Non-limiting examples of suitable dosage forms include tablets, capsules or sachets. Since certain digestive enzymes, such as pancreatic lipases, need to be protected from gastric inactivation prior to release into the duodenum, it may also be desirable that the stabilized digestive enzyme compositions or oral dosage forms of the present invention be provided as a release formulation controlled or delayed. Such controlled- or delayed-release formulations can include tablets or particles coated with an enteric coating that serves to protect pH-sensitive digestive enzymes from gastric inactivation while still releasing the digestive enzymes into the duodenum. Alternatively, controlled release formulations can include capsules filled with the stabilized digestive enzyme compositions or oral dosage forms of the present invention, whereby the capsule protects the contents against gastric inactivation, yet releases digestive enzymes into the duodenum. However, the stabilized digestive enzyme compositions or oral dosage forms of the present invention are not limited to digestive enzymes susceptible to gastric inactivation, for example, certain digestive enzymes that are naturally stable in the gastric environment such as gastric lipases, a range of proteases , including those of pancreatic origin and amylases. Certain digestive enzymes derived or extracted from microorganisms that have intrinsic stability or that have been chemically modified through cross-linking.

When the compositions of the present invention are formulated as tablets, the stabilized digestive enzyme(s) may be "compressed" (i.e., shaped into tablets) using methods known in the art and subsequently coated with a enteric coating, again using methods known in the art.

When the compositions of the present invention are formulated as capsules, the capsule contents can be formulated using methods known in the art. For example, the stabilized digestive enzyme composition can be provided in the form of particles or tablets suitable for incorporation into a capsule.

The term "particles" as used herein includes fine powders (having particle diameters in the range of about 1 µm) to pellets having a diameter of about 5 mm.

The stabilized digestive enzyme composition can also be formed into particles coated with a coating, where the coating comprises an enteric polymer. The term "enteric polymer" means a polymer that protects digestive enzymes from gastric contents, for example, a polymer that is stable at acidic pH but can break down quickly at higher pH or a polymer whose range of hydration or erosion is slow enough to ensure that contact of gastric contents with digestive enzymes is relatively small while it is in the stomach, as opposed to the rest of the gastrointestinal tract. Non-limiting examples of enteric polymers include those known in the art, such as modified or unmodified natural polymers such as cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, and shellac; or synthetic polymers such as acrylic polymers or copolymers of acrylic acid polymers and copolymers, methylmethacrylate copolymers and methacrylic acid/methylmethacrylate copolymers.

The enteric polymer coating can be a synthetic polymer, optionally including an organic material such as an alkalizing agent. The resulting coated particles provide delayed release beads comprising a core comprising the stabilized digestive enzyme(s) and an enteric coating encapsulating the core. The coated stabilized digestive enzyme particles can then be formulated into tablets or capsules.

The enteric polymer and the at least one inorganic material impart enteric properties to the coating of the present invention. That is, when used as a medication, the coating will act as a barrier protecting the medication from the acidic environment of the stomach and substantially prevent the release of the medication before it reaches the small intestine (ie, the release of enzymes in the stomach is less than that about 10-20% of the total amount of enzyme in the composition).

The inorganic material can include, for example, an alkalizing agent. Non-limiting examples of alkalizing agents include silicon dioxide, sodium salts, calcium salts, magnesium salts, aluminum salts, aluminum hydroxides, calcium hydroxides, magnesium hydroxide, talc and combinations thereof. In one embodiment, the alkalizing agent is talc.

Depending on the intended use of the composition, the ratio of the enteric polymer and the at least one inorganic material can range from about 10:1 to about 1:60 by weight. In another embodiment, the ratio of enteric polymer and the at least one inorganic material ranges from about 8:1 to about 1:50 by weight. In another embodiment, the ratio of the enteric polymer and the at least one inorganic material ranges from about 6:1 to about 1:40 by weight. In another embodiment, the ratio of the enteric polymer and the at least one inorganic material ranges from about 5:1 to about 1:30 by weight. In another embodiment, the ratio of the enteric polymer and the at least one inorganic material ranges from about 4:1 to about 1:25 by weight. In another embodiment, the ratio of the enteric polymer and the at least one inorganic material ranges from about 4:1 to about 1:9 by weight. In another embodiment, the ratio of the enteric polymer and the at least one inorganic material ranges from about 10:4 to about 10:7 by weight.

In one embodiment, the compositions or oral dosage forms of the present invention comprise particles of stabilized digestive enzyme coated with an enteric coating comprising an enteric polymer and an inorganic material such as talc. In a particular embodiment, the inorganic enteric coating material comprises about 1-10% by weight of the total weight of the particles. In another embodiment, the inorganic material comprises about 3, about 5, about 7 or about 10% by weight of the particles. In still other embodiments, the inorganic material is an alkalizing agent and comprises about 20-60% by weight of the dry coating. In still other embodiments, the alkalizing agent is about 25%, about 30%, about 35%, about 40%, about 45%, about 50% or about 55% of the dry coating weight (including all ranges, sub-ranges and values in between). In a particular embodiment, the alkalizing agent is talc. In yet another particular embodiment, the dry coating of the particles comprises about 35% talc.

In another embodiment of the present invention, the coating further comprises a plasticizer. Examples of suitable plasticizers include, but are not limited to, triacetin, tributyl citrate, triethyl citrate, acetyl tri-n-butyl citrate, diethyl phthalate, dibutyl sebacate, polyethylene glycol, polypropylene glycol, castor oil , acetylated monoglyceride, acetylated diglyceride and mixtures thereof.

The dosage forms of the present invention can be capsules containing the composition of the present invention (e.g., controlled release particles of the stabilized digestive enzyme composition, coated with an enteric polymer and an alkalizing agent). The capsules themselves can be comprised of any conventional biodegradable material known in the art, for example, gelatin, polysaccharides such as pullulan or modified cellulosic materials such as hydroxypropylmethylcellulose. In order to improve the stability of the stabilized digestive enzymes, capsules can be dried before filling, or a capsule comprised of a low moisture content material can be selected. In one embodiment of the dosage form of the present invention, the capsule is comprised of hydroxypropylmethylcellulose. In another embodiment of the dosage form of the present invention, the capsule is comprised of hydroxypropylmethylcellulose having a water content of about 6% or less, for example, about any of 4% or less, 2% or less, or 2- 6% or 4-6%. In another embodiment, the capsule is comprised of hydroxypropylmethylcellulose having a water content of less than about 2%.

The dosage forms of the present invention can comprise a simple digestive enzyme or mixtures of digestive enzymes. If the stabilized digestive enzyme composition is formed into particles coated with an enteric coating, the coated particles can each contain a core comprising a simple digestive enzyme or digestive enzyme mixtures. The dosage form can also comprise coated particles, each of which is nominally the same composition, or it can comprise mixtures of different types of coated particles. For example, the dosage form can be a capsule filled with coated particles, where each of the coated particles has a core comprising pancrelipase. Alternatively, the dosage form can be a capsule filled with coated particles, where some of the coated particles have a core comprising pancrelipase, while other coated particles have cores comprising a different lipase, or proteases or amylases. Any suitable combination of coated particles of different compositions can be used to provide the desired therapeutic effect.

Furthermore, when dosage forms of the present invention are comprised of stabilized digestive enzyme coated particles, the individual particles may each have the same coating composition or may include mixtures of particles, some of which have a different coating composition. Any suitable combination of coating compositions can be used to provide the desired type of controlled release or therapeutic effect.

The core of the coated particles can be of any suitable particle size or shape. For example, the coated particles may be in the form of a coated powder having a particle size range of about 50-5000 microns or may be in the form of "minitabs" having a nominal particle diameter in the range of about 2 -5 mm. For other applications, the core of the coated particles may be "microtabs" which have nominal particle diameters of less than about 2 mm, for example about 1-2 mm.

In one embodiment, the compositions or oral dosage forms of the present invention can comprise a plurality of coated digestive enzyme particles (e.g., pancrelipase). The digestive enzyme particles can comprise a digestive enzyme, at least one disintegrant, at least one polymeric binder or diluent and optionally at least one plasticizer, optionally at least one anti-adherent and optionally at least one lubricant. In one embodiment, the digestive enzyme particles can comprise about 60-90% digestive enzyme, about 1-4% of at least one disintegrant, about 2-6% of at least one polymeric binder or diluent, and optionally about 0.5-1.0% of at least one plasticizer, optionally about 0.2-0.6% of at least one non-stick, and optionally about 0.2-0.6% of at least one lubricant. For example, the digestive enzyme particles can comprise about 60-90% pancrelipase, about 1-4% croscarmellose sodium, about 0.5-1.0% hydrogenated castor oil, about 0. 2-0.6% silicon dioxide, about 2-6% microcrystalline cellulose and about 0.2-0.6% magnesium stearate. The coating can comprise at least one enteric polymer, about 4-10% of at least one alkalizing agent (based on the total weight of the particles) and optionally at least one plasticizer. In one embodiment, the coating can comprise about 10-20% of at least one enteric polymer, about 4-10% of at least one alkalizing agent, and about 1-2% of at least one plasticizer (based on weight total particle size). For example, the coating can comprise about 10-20% hydroxypropylmethylcellulose phthalate, about 4-10% talc and about 1-2% triethyl acetate (based on the total weight of the particles). The plurality of coated digestive enzyme particles can then be formed into a tablet or filled into a capsule. In one embodiment, the capsule comprises hydroxypropylmethylcellulose.

The compositions of the present invention and dosage forms comprising the compositions of the present invention have improved stability compared to conventional digestive enzyme compositions and dosage forms (e.g., pancrelipase). Consequently, the dosage forms of the present invention do not require "overfilling" (i.e., zero overfilling), as do conventional digestive enzyme dosage forms, to deliver a clinically useful amount of digestive enzyme to a patient. in need of it. Conventional digestive enzyme compositions and dosage forms require fill levels in excess of up to 65% (i.e., 165% of the required dose of digestive enzyme) to compensate for poor enzyme stability. As a result, there is uncertainty of the dose applied by conventional digestive enzyme compositions. Thus, "over-filled" dosage forms may apply more than the intended dose of digestive enzymes shortly after manufacture, but over time, enzyme activity may drop below the intended dose.

In one embodiment, the dosage forms comprising the compositions of the present invention are substantially zero excess fill. The term "substantially zero excess filler" means compositions of the present invention where the amount of additional digestive enzyme activity (i.e., the amount of additional enzyme activity above the target dose) is less than or equal to about 10 %, that is, about 10%, less than about 10%, less than or equal to 9%, less than or equal to 8%, less than or equal to about 7%, less than or equal to about 6%, less than or equal to about 5%, less than or equal to about 4%, less than or equal to about 3%, less than or equal to about 2 %, less than or equal to about 1% or about 0%. So, for example, if the desired dose is about 4500 IU of lipase, the substantially zero excess fill dosage forms of the present invention may contain less than or equal to about 4950 IU of lipase (i.e., less than that or equal to 110% of 4500 IU of lipase). In another embodiment, the zero overfill dosage form contains 4500 IU of lipase.

Compositions or dosage forms (for example, tablets or capsules) of the present invention may be stored in any suitable packaging. For example, the packaging could be a glass or plastic jar with a screw or snap closure. Alternatively, the compositions or dosage forms of the present invention can be packaged as a unit dosage form in blister packs. Applicant has found that improved stability of digestive enzyme compositions or dosage forms can be provided by providing a moisture-proof seal and/or moisture-proof packaging. Non-limiting examples of moisture-proof packaging include glass jars, plastic jars incorporating moisture barrier resins or coatings, aluminized plastic packaging (eg Mylar), etc. The term "moisture proof" refers to a package that has a water permeability of less than about 0.5 mg of water per cm3 of container volume per year.

Containers (eg bottles) can be closed with any suitable closure, especially closures that minimize moisture ingress during storage. For example, the compositions or dosage forms of the present invention can be closed with a closure such as Saf-Cap III-A (Van Blarcom Closures, Inc.) containing HS 035 Heat Seal/20F (SANCAP Liner Technology, Inc.) , printed as a sealing line.

In order to ensure package integrity and minimize moisture ingress during storage, sealed packages containing the compositions or dosage forms of the present invention may be leak tested after applying the composition or dosage form of the present invention and sealing the package. For example, sealed packages can be tested by applying a controlled vacuum to the closure, and detecting a decrease in vacuum over time. Suitable leak testing equipment includes those manufactured by Bonfiglioli (eg model LF-01-PK or model PKV 516).

Packages containing the compositions or dosage forms of the present invention may also contain a desiccant (i.e., a substance that absorbs, reacts with or adsorbs water) capable of reducing moisture within the package, e.g., a desiccant capsule, capable of " sequestering" moisture from a sealed atmosphere within the package. Non-limiting examples of suitable desiccants that can be placed within such packages include zeolites (eg molecular sieves such as 4A molecular sieves), clay (eg montmorillonite clay), silica gel, activated carbon or combinations thereof . In one embodiment, the desiccant comprises molecular sieves.

Furthermore, it is common practice when packaging oral pharmaceutical unit doses to add a "buffer" of a cellulosic material, such as cotton, into the top of the container to fill the void space in the top of the container, thereby minimizing movement of contents.

Cellulosic materials are somewhat hygroscopic, and can act as a "reservoir" of moisture within the package. Thus, in one embodiment of the packaging of the present invention, no cellulosic or cotton "buffer" is present in the packaging. In another embodiment of the packaging of the present invention, the packaging does not have a cellulosic or cotton plug and contains a desiccant.

Compositions of the present invention can be prepared using conventional techniques, but modified as indicated herein to provide moisture contents of about 3% or less, water activities of about 0.6 or less, or provide stabilized digestive enzyme compositions. which exhibit an activity loss of no more than about 15% after a three-month accelerated stability test. For example, digestive enzyme particles (eg pancrelipase) can be re-coated in a fluid bed coater equipped with a dehumidifier. In one embodiment, the coating apparatus is operated in an atmosphere having a water content of about 4 g/m3 or less, about 3.5 g/m3 or less, about 3 g/m3 or less, about 2.5 g/m3 or less, about 2.0 g/m3 or less, about 1.5 g/m3 or less, about 1.0 g/m3 or less, or about 0.5 g/m3 or less, including all tracks and subtracks in between. The atmosphere where the coating is carried out may comprise dehumidified air, dehumidified nitrogen or other dehumidified inert gas.

The coating can be applied as a solution of the enteric polymer (and optionally a suspended organic material) in an organic solvent such as an alcohol (eg ethanol), a ketone (eg acetone), methylene chloride or mixtures thereof (eg acetone/ethanol mixtures).

The compositions of the present invention provide improved absorption of fats, proteins and carbohydrates in patients suffering from conditions or disorders associated with a digestive enzyme deficiency. In one embodiment, compositions of the invention, in particular pancrelipase or pancreatin compositions, can be used to treat exocrine pancreatic insufficiency (EPI) associated with various diseases. Such diseases include, but are not limited to, cystic fibrosis (CF). In some embodiments, such compositions can substantially alleviate malabsorption (e.g., from fats) associated with EPI in cystic fibrosis patients and other patients, including pediatric patients. In some embodiments, such compositions can increase the fat absorption coefficient (CFA) - to at least about 85% or more in patients with cystic fibrosis. Such results can be achieved when co-administered with other agents or compositions or can be achieved without co-administration with other agents. In one embodiment, such CFA results are achieved without co-administration of proton pump inhibitors such as Prilosec®, Nexium® and the like.

For patients identified as having low GL pH levels (e.g., GL pH levels < than about 4), improved results can be obtained by administering the compositions or dosage forms of the present invention together with pump inhibitors of proton, antacids and other drugs that increase the pH of the Gl tract. For example, the compositions or dosage forms of the present invention can be administered separately from proton pump inhibitors, antacid or other drugs (or prior to, concomitantly with or after administration of proton pump inhibitor, antacid, etc). Alternatively, the proton pump inhibitor, antacid or other drug can be combined with the pancreatin composition of the present invention as a single dosage form.

In yet another embodiment, the present invention provides a method of treating or preventing a disorder associated with a digestive enzyme deficiency comprising administering a composition of the present invention to a mammal in need thereof. In one embodiment, the mammal is a human.

In yet another embodiment, the present invention provides a method of treating or preventing a disorder associated with deficiency of a digestive enzyme comprising administering a composition or dosage form of the present invention to a mammal in need thereof, wherein the composition or dosage form of the present invention comprises, in addition to at least one digestive enzyme, a proton pump inhibitor, antacid, or other drug which increases the pH of Gl. In yet another embodiment, the present invention provides a method of treatment or preventing a disorder associated with deficiency of a digestive enzyme comprising administering a composition or dosage form of the present invention in combination with a dosage form comprising a proton pump inhibitor, antacid or other drug that increases the pH of G1.

Disorders that can be treated with the composition or dosage form of the present invention include conditions where the patient does not have any or low levels of digestive enzymes or where patients require digestive enzyme supplementation. For example, such conditions can include cystic fibrosis, chronic pancreatitis, other pancreatic diseases (eg, hereditary, post-traumatic and allograft pancreatitis, hemochromatosis, Shwachman syndrome, lipomatosis or hyperthyroidism), side effects of cancer or cancer treatment, side effects of surgery (eg, gastrointestinal bypass surgery, Whipple procedure, total pancreatectomy, etc.) or other conditions where pancreatic enzymes cannot reach the intestine, poor mixing (eg, Billroth II gastrectomy, other types of surgery of gastric bypass, gastronomy, etc.), side effects of drug treatments such as metformin treatment or those drugs used to treat symptoms of HIV and autoimmune diseases such as diabetes where the pancreas may be compromised, obstruction (eg, pancreatic and biliary duct lithiasis, pancreatic and duodenal neoplasms, ductal stenosis), malabsorption associated with disease celiac disease, food allergies and aging.

The amount of the composition or dosage form of the present invention administered daily to mammals (e.g., humans) depends on the desired result. The knowledgeable physician will be able to prescribe the required dose based on their diagnosis of the condition to be treated.

For example, for the treatment of digestive enzyme insufficiency in humans (eg related to cystic fibrosis) the starting dose should be 500 to 1000 lipase units/kg/meal, with the total dose not exceeding 2500 lipase units/kg /meal or 4000 units of lipase/g fat/meal as per US FDA recommendations. Typically, a patient should receive at least 4 dosage forms per day, preferably given with food.

Examples Example 1

*Método USPPancrelipase MT (mini-tablets) is a mixture of pancrelipase raw material (eg, obtained from Nordmark) and excipients in tablet form using 2 mm diameter beveled perforations. The physical characteristics of Pancrelipase MT before coating are shown below in Table 1.

*USP method

Pancrelipase MT was coated with a coating formulation (Table 2) using a Glatt-GPCG1 fluid bed apparatus equipped with a Munters ML 1350 dehumidifier in the process air flow. The coating process was carried out with process air at three different moisture contents (Table 3). For each batch, the coating weight was approximately 15% of the total weight of the coated particles. The composition of the coated particles for each set of process conditions is approximately the same (Table 4), and appeared uniform, smooth and homogeneous upon microscopic examination.

The three sets of samples (ie, P9A165, P9A167 and 5 pP9A170) showed residual moisture contents corresponding to the moisture content of the process air stream (Table 4).

The influence of residual moisture on activity loss with time was evaluated under accelerated stability conditions as follows: Hard gelatin capsules (dosage 20,000 IU lipase) were filled with the three batches of coated Pancrelipase MT mini-tablets described above and stored at 40°C at 75% relative humidity in sealed Nialene bags. Lipase activity was evaluated after 15 days and 4 months of storage. The results are shown in Table 6.

The results in Table 6 show that improved stability is provided by compositions having a moisture content of less than about 2%. Alternatively, improved stability is provided by coating under an atmosphere with a moisture content of less than 3.6 g/m3 to 0.4 g/m3.

Example 2

Pancrelipase MT particles were coated with two coating compositions containing different amounts of talc (Table 7) Table 7

Coating tests were carried out using a Glatt-GPCG1 fluid bed apparatus equipped with a Munters ML 1350 dehumidifier in order to ensure process airflow at a low moisture content (ie less than 1 g/m3). Coating weights were approximately 15%. The theoretical composition of the batches is reported in Table 8. Microscopic examination indicated that the coatings on all samples were smooth and homogeneous. Residual moisture contents were measured by loss on drying (Table 9).

The effects of different coating compositions on activity loss over time were evaluated under accelerated stability conditions as follows: Hard gelatin capsules (dosage 20,000 Lipase III) were filled with the two batches of coated Pancrelipase MT described above and 15 stored at 40°C and 75% relative humidity in sealed Nialene bags.

meses de armazenamento sob condições de estabilidade acelerada é signi- ficantemente menor para amostras revestidas com um revestimento com alto teor de talco (Lote P9A240). Deste modo, aumento da concentração de talco 5 de a partir de aproximadamente 1 % para aproximadamente 7% resulta em aperfeiçoamentos significantes em estabilidade de enzima.Lipase activity was checked after 1, 2 and 3 months of storage as shown in Table 9.

months of storage under accelerated stability conditions is significantly less for samples coated with a high talc coating (Lot P9A240). Thus, increasing the talc 5 concentration from approximately 1% to approximately 7% results in significant improvements in enzyme stability.

Example 3

Glatt-GPCG1 de leito fluidizado equipado com um desumidificador Munters ML 1350 a fim de assegurar fluxo de ar de processo em um teor de umidade baixa (menor do que 1 g/m3). Os pesos dos revestimentos eram aproximadamente 15%. A composição teórica das duas bateladas é relatada na Tabe- 5 la 12. Tabela 12

The solvent effects of coating composition were evaluated by preparing "high talc" and "low talc" coating compositions similar to those described in Table 6, except that the solvent ethanol (96% ethanol, 4% of water)/acetone was replaced with 100% acetone (Table 11).

Glatt-GPCG1 fluid bed equipped with a Munters ML 1350 dehumidifier in order to ensure process air flow at a low moisture content (less than 1 g/m3). Coating weights were approximately 15%. The theoretical composition of the two batches is reported in Table 5 12. Table 12

Lot P9A318 complies with commercial product specifications, but Lot P9A352 did not pass the gastro-resistance test. Microscopic examination showed that the film coating of Lot P9A352 was not as smooth and homogeneous as the other coated samples, probably because of the higher evaporation rate of acetone compared to the ethanol/acetone mixture used in previous samples. and the high concentration of talc in the coating.

comparado com a estabilidade do Lote P9A230, que foi preparado com um revestimento similar sob condições de revestimento similares (Tabela 14).Lot P9A318 was then evaluated under accelerated stability conditions as follows: Hard gelatine capsules (dosage of 20,000 IU Lipase) were prepared and stored at 40°C and 75% relative humidity in sealed Nialene pouches. Lipase activity was measured after 1, 2 and 3 months of storage as shown in Table 13. Table 13

compared to the stability of Lot P9A230, which was prepared with a similar coating under similar coating conditions (Table 14).

It appears then that replacing 96% ethanol with acetone in the coating formulation provides significantly less loss of enzyme activity over time. Table 14

Example 4

Conforme mostrado nas tabelas 15 a 17. composições de lipase em cápsulas de HPMC mostram atividade de lipase significantemente maior após armazenamento por 15, 30 e 90 dias sob condições de estabilidade acelerada e cápsulas de HPMC secas oferecem melhor estabilidade do que aquelas que contêm níveis de umidade em equilíbrio.CPS and HPMC (hydroxypropyl methylcellulose) gelatin capsules were filled with identical coated lipase compositions. The water content of gelatine capsules is approximately 14% and the water content of 10 HPMC capsules is approximately 4%. Yet a set of HPMC capsules were dried to a moisture level of less than 2%. All samples were subjected to accelerated stability conditions (40°C and 75% relative humidity; heat sealed samples in Nialene bags and lipase activity were tested after 15, 30 and 90 days. The results are shown below in the tables 15 to 17. 1) HPMC CPS vs GELATIN CPS

As shown in tables 15 to 17, lipase compositions in HPMC capsules show significantly higher lipase activity after storage for 15, 30 and 90 days under accelerated stability conditions and dry HPMC capsules offer better stability than those containing levels of moisture in balance.

Example 5

Gelatin and hydroxypropylmethylcellulose capsules were filled with coated lipase compositions in Mini-tablet form. The coating for the gelatin capsule compositions (P200050) contained approximately 10% talc, while the coating for the hydroxypropylmethylcellulose capsule compositions (P200550) contained approximately 33% talc. The coating compositions were otherwise identical. Table 18 below compares the levels of degradation observed after storage under conditions of accelerated stability with the moisture content of the compositions. As shown in Table 18, higher levels of lipase activity are related to lower levels of moisture in the composition. Furthermore, compositions filled in HPMC capsules are more stable than compositions filled in gelatine capsules. Table 18

Example 6

The storage effects of capsules containing lipase compositions in packages containing a desiccant were evaluated by measuring the lipase activity in the samples after 30 and 90 days of storage under accelerated stability conditions (40°C and 75% relative humidity; heat sealed samples in Nialene pouches). As shown in Tables 19 and 20, lipase activity is significantly higher in packages containing a desiccant and in capsules that are dried below equilibrium moisture content. 2) DISSECANTS Desiccant 1: silica gel in Tyvek® bags Desiccant 2: molecular sieves in Tyvek® bags

Example 7

Pancrelipase MT particles were coated with two coating compositions having a talc level intermediate between the "low" and "high" levels employed above (HP55:TEC:Talco=10:1:5), using either acetone or a mixture of ethanol/acetone as the coating solvent. The theoretical composition of the two coating suspensions shown in table 21 below. Continuation...

Coating tests were carried out using an apparatus

Glatt-GPCG1 fluid bed equipped with a Munters ML 1350 dehumidifier in order to ensure process air flow at a low moisture content (less than 1 g/m3).

The batches were prepared by coating Pancrelipase MT in a coating weight of approximately 15%. Three batches were prepared with an ethanol/acetone coating solvent and the three batches were prepared with an acetone coating solvent. The theoretical composition, which was the same for all six batches, is shown below in table 22. Table 22

Microscopic examination of the coating for all six samples appeared smooth and homogeneous. The coated Pancrelipase MT particles were then filled into HPMC capsules and packaged in glass bottles containing desiccants (molecular sieves). The bottles were then sealed, stored under conditions of accelerated stability and lipase activity was evaluated at various time periods as indicated below in table 23.

The packaging conditions for each sample were as follows. Twelve HPMC capsules (strength 20,000 IU Lipase) and 1 g of 10 molecular sieves (Minipax-Multisorb absorbent) as desiccant were placed in a 30 ml glass bottle. The bottles were capped with Saf-Cap III-A containing HS 035 Heat Seal/20F printed as a seal liner and stored at 40°C/75% RH.

As shown in table 23, the three samples prepared with the ethanol/acetone coating solvent showed similar losses in lipase activity. After 2 months of storage, two of the samples prepared with the acetone coating solvent showed no loss of activity, and the third showed an activity reduction of 4%. This suggests that samples prepared with the acetone coating solvent are more stable than samples prepared with the ethanol/acetone coating solvent.

Example 8

Microtablets To provide additional choices for dosage formulations were made where the tablet dimensions were significantly reduced. The pancrelipase mixture was formed into a tablet with a diameter of 1.5 mm in circumference, radius of 1.2 mm and perforations of 15 curvature. Compression parameters were adjusted to obtain microtablets ("μT") with friability of less than 2.5% (USP method). The characteristics of Lot 9A402 are shown in table 24,

Batch P9A402 was coated in a Glatt-GPCG1 fluid bed apparatus equipped with a Munters ML 1350 dehumidifier in order to ensure process air flow at low moisture content (less than 15 g/m3) with a suspension having the composition shown in table 2. A coating weight of 22% was obtained. Microscopic examination of the film coatings indicated that all samples appeared smooth and homogeneous.

The theoretical composition of Batch P9A422 is shown 10 in table 25. Table 25

Two other batches of microtablets were prepared as described above and their properties are shown below in Table 26.

Pancrelipase microtablets were coated with one of two suspensions having talc intermediate levels between the "high" and "low" levels described above (HP55:TEC:Talco=10:1:5), using either acetone or a mixture of ethanol in acetone as a coating solvent (table 5 to 27).

The six tests were performed using a Glatt-

As composições teóricas das bateladas são sumarizadas na ta bela 28.

Fluid bed GPCG1 equipped with a Munters ML 1350 dehumidifier to ensure process airflow at low moisture content (less than 1 g/m3). Coating weights were approximately 10 22% and microscopic examination indicated that the coatings were smooth and homogeneous.

The theoretical compositions of the batches are summarized in table 28.

HPMC cps capsules were filled with the coated microtablets described above and packaged in glass bottles containing desiccants (molecular sieves). The bottles were then sealed with Saf-Cap III-A, containing HS 035 Heat Seal/20F printed as a seal aligner and stored under accelerated stability conditions (40°C and 75% relative humidity). Twelve MPMC capsules (strength 5,000 IU Lipase) and 1 g of molecular sieves (Minipax-Multisorb absorbent) as desiccant were placed in a 30 ml capacity glass bottle. Lipase activity was measured at 20, 30, 40 and 60 days of storage as shown in tables 29 and 30.

All three samples prepared using an ethanol/acetone coating solvent showed similar behavior and after months of storage, the loss of lipase activity was 2% to 4%. After two months of storage the loss of activity of two of the samples prepared using an acetone coating solvent showed no evidence of loss of lipase activity, while a third sample showed a 5% decrease in lipase activity. Thus, compositions prepared with acetone as the coating solvent were more stable than samples prepared with ethanol/acetone coating solvent, possibly linked to the water content of the ethanol used.

The microtablets prepared above were slightly oblong (see tables 24 and 26); the thickness to diameter ratio of the microtablets was between 1.22:1 and 1.15:1.

To further reduce the dimensions of the microtablets, new samples that were prepared with thickness to diameter ratios of almost 1:1 (Lot Q9A006) are shown below in Table 31.

Lot Q9A006 was coated with the compositions shown in table 32 at a coating weight of 22%. The coating tests were carried out using a Glatt-GPCG1 fluid bed "eyes" apparatus equipped with a Munters ML 1350 dehumidifier in order to ensure airflow processing at low moisture content (less than 120 g/m3).

The theoretical composition of coated microtablet Lot Q9A019 was the same as that shown in table 28. Microscopic examination indicated that the coatings were smooth and homogeneous.

The above examples show that digestive enzyme compositions with improved stability can be prepared by maintaining low moisture contents and water activities in the composition components, for example, replacing aqueous ethanol/acetone coating solvents with acetone, coating coating mini-tablets and micro-tablets in dehumidified process air streams (eg having moisture contents between 0.4 g/m3 and 3.6 g/m3). Also, high levels of inorganic materials in the coating (eg HP55:TEC:Talc ratios ranging from 10:1:1 to 10:1:5), selection of less hygroscopic capsule materials (eg HPMC or Dry HPMC) and improved packaging techniques (eg, storage in tightly sealed glass bottles containing desiccants) provide digestive enzyme compositions and dosage forms with improved stability.

Example 9

Table 33 below shows accelerated stability testing (in bottles; 40°C and 75% relative humidity) of pancrelipase mini-tablets coated with Eudragit. Table 33

The results indicate that conventional enteric coatings such as Eudragit do not provide stabilized pancrelipase compositions.

Example 10

Examples of dosage forms comprising variable dosage ER coated beads per capsule, coated as described in the previous examples, are shown in Table 34, below: Table 34

Example 11

Table 35 below shows the water content of containers of various sizes containing capsules comprising the compositions of the present invention. The water content includes total water from the capsules and water permeating into the container for a storage time of two years. The "weight of equivalent molecular sieves" is the minimum amount of molecular sieves required to absorb the water present in the containers. Table 35

Example 12

A double-blind, randomized, placebo-controlled, phase III crossover study was performed to compare the effects of treatment with the Tabeela 34 pancrelipase compositions with those of a piacebo in 34 seven-year-old and 34 CF patients with EPI. Older. The study was conducted at 14 CF centers across the US. The main endpoint of the study compared the fat absorption coefficient following oral administration of the pancrelipase composition at daily doses of less than or equal to 10,000 lipase units per kilogram of body weight in combinations of 5,000, 10,000, 15,000 or 20,000 lipase units per capsule versus placebo. The secondary endpoints of the test evaluated changes in the nitrogen absorption coefficient as a measure of protein absorption, cholesterol, fat-soluble vitamins, weight, body mass index, and EPI symptoms.

Patients treated with these compositions showed a statistically significant increase in fat absorption coefficient and nitrogen absorption coefficient compared to those receiving a placebo and had fewer symptoms associated with impaired absorption such as bloating, flatulence, pain and evidence of fat in the stool. . Increases in mean cholesterol and vitamin levels were also observed in patients taking these pancrelipase compositions versus placebo. A statistically significant decrease was obtained in the frequency of stools per day. These compositions were well tolerated by patients, and any serious drug-related adverse events were noted during the study.

The mean percentage coefficient of fat absorption in patients receiving these compositions was 88.28% versus 62.76% in patients receiving placebo. The mean percentage coefficient of nitrogen absorption was 87.2% versus 65.7% in patients taking placebo and the number of stools per day decreased in respective patient groups from 2.66 to 1.77.

Example 13

A Pediatric Phase III clinical trial was performed to evaluate the effects of treatment with the compositions in Table 34 in an open-label study in 19 CF patients less than seven years old at 11 CF treatment centers in the US - the first trial of pancreatic replacement therapy of this size conducted in young children and infants with exocrine pancreatic insufficiency. Study design involved a seven-day dose stabilization period followed by a seven-day treatment period; patients received 5,000 units of lipase per capsule daily, with the product being spread over food as needed. The primary endpoint of the study was the percentage of "responders" or those patients with excess stool fat and no signs and symptoms of malabsorption after one and two weeks of treatment. Secondary endpoints included weight change, nutritional status, stool frequency and consistency, incidences of bloating, pain and flatulence as well as physician and parent or guardian judgment of improved clinical symptoms. Product safety was also evaluated.

The mean percentage of responders, as defined in the study protocol, at assessment (the start of the stabilization period when patients were on prior pancreatic enzyme replacement therapy and before treatment) was 52.6%. At the end of the stabilization period and at the end of the treatment period, the mean percentages of responders were 66.4% and 57.9%, respectively. Among children in the study, symptoms of malabsorption were significantly less at the end of the treatment period than at screening, in accordance with observations regarding control of malabsorption symptoms seen in the Phase III trial described in Example 12, above. The pancrelipase compositions of the present invention were also well tolerated by these patients and any serious drug-related adverse events were observed during the study.

The results showed that the compositions of the present invention effectively controlled the signs and symptoms of malabsorption and support the results obtained in the pivotal phase III trial described in Example 12. A significant proportion of physicians and patients felt that symptom control with the compositions of the present invention improved versus previous therapies.

Example 14

An open-label, Phase III, randomized, single-center, single-treatment, crossover study was performed to compare treatment effects with the pancrelipase compositions in Table 34 to determine the gastric bioavailability of these compositions under feeding conditions in 10 patients with chronic pancreatitis with exocrine pancreatic insufficiency. Exclusionary drugs (proton pump inhibitors (PPI's), antacids and drugs capable of altering Gl mobility were discontinued 7 days before entering the study. Patients were randomized to receive or Ensure Plus® (a nutritional supplement fortified with vitamin available from Abbott) alone or Ensure Plus® in combination with 75,000 units of USP lipase (3 capsules containing 20,000 units each plus 3 capsules containing 5000 units each) per procedure.The capsules were opened and their contents mixed with 480 ml_ Ensure Plus ® immediately prior to administration. After a one-day washout period, this procedure was repeated, except that patients who previously received Ensure Plus® alone received Ensure Plus® in combination with 75,000 units of USP lipase and patients who previously received the combination of Ensure Plus® and lipase received Ensure Plus® alone. The next day, patients received a physical examination, and blood and urine samples were collected. The bioavailability of the compositions of the present invention was estimated from the amount of the respective enzymes released and recovered (i.e., lipase, amylase and chymotrypsin) in the duodenum following administration of the composition in the presence of Ensure Plus®. Measurements of blood cholecystokinin levels and gastric and duodenal pHs were also performed. Lipase activity was measured according to the method of Carriere, F.; Barrowman, J.A.; Verger, R.; Laugier, R. "Secretion and contribution of lipolysis of gastric and pancreatic lipases during a test meal in humans". Gastroenterology 1993, 105, 876-88. Amylase and chymotrypsin were measured according to the methods described in Carriere, F.; Grandval, P.; Renou, C.; Palomba, A.; Prieri, F.; Giallo, J.; Henninges, F.; Sander-Struckmeirs, S.; Laugier, R. "Quantitative study of digestive enzyme secretion and gastrointestinal lipolysis in chronic pancreatitis". Clin. Gastroenterol. Hepatol. 2005, 3, 28-38.

Treatment with the pancrelipase compositions of the present invention was found to result in statistically significantly higher amounts of amylase, lipase and chymotrypsin released in the duodenum of patients who received the combination of Ensure Plus® and pancrelipase, compared to patients who received Ensure Plus® alone (after pH correction as a confounding factor).

The mean bioavailability for lipase, amylase and chymotrypsin for the eight patients who completed the study according to the protocol was 27.5%, 21.6% and 40.1%, respectively. The patient was found to fit into two different GL pH subpopulations ("normal pH" and "low pH"). For patients who had "normal pH" values (ie, patients with a mean duodenal pH greater than 4), the mean bioavailability of lipase, amylase and chymotrypsin was greater than for the entire study group: 45.6% , 26.9% and 47.7%, respectively. No difference in cholecystokinin values between the two treatments was observed.

Because the bioavailability for lipase, amylase and chymotrypsin differs for "normal pH" and "low pH" patients (particularly for lipase), the effectiveness of pancrelipase compositions or dosage forms of the present invention may be increased, for example, in patients with "low pH" by co-administration of drugs that increase the pH of GI, e.g., PPI's and antacids. However, the compositions or dosage forms of the present invention may be administered without co-administration of, e.g., PPI's .

The above description of the invention has been presented for the purpose of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form described. Modifications and variations are possible in light of the above teachings. The descriptions of the embodiments have been chosen in order to explain and describe the principles of the present invention and its practical application and are not intended to limit the scope of the claims.

All publications and patents or patent applications mentioned herein are incorporated by reference in their entirety to the extent as if each individual publication, patent or patent application were specifically and individually indicated and incorporated by reference.

Claims (42)

1. Composition, characterized in that it comprises at least one digestive enzyme as a plurality of coated digestive enzyme particles, wherein the at least one digestive enzyme is pancrelipase, wherein each said particle contains a core comprising pancrelipase coated with a coating enteric, and, that the enteric coating comprises an enteric polymer and further comprises 4 to 10% talc based on the total particle weight, wherein the composition has a moisture content of 3% or less. 2. Composition according to claim 1, characterized in that the core is a "minitab" having a diameter of 2 to 5 mm. 3. Composition according to claim 1, characterized in that the core is a "microtab" having a diameter of 1 to 2 mm. 4. Composition according to claim 1, characterized in that the composition has a moisture content between 1.5% and 2.5%. 5. Composition according to claim 1, characterized in that the composition has a moisture content of 2% or less. 6. Composition according to claim 1, characterized in that the composition has a moisture content of 2%. 7. Composition according to claim 1, characterized in that it comprises at least one pharmaceutically acceptable excipient. 8. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is selected from the group consisting of a binder, disintegrant, lubricant, non-stick, diluents and mixtures thereof. 9. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one stabilizer selected from the group consisting of trehalose, proline, dextran, maltose, sucrose, mannitol, polyols, silica gel , aminoquanidine, pyridozamine and mixtures thereof. 9. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one binder selected from the group consisting of starches, sugars, lactose, their sugar alcohols, a xylitol, a sorbitol, maltitol , cellulose, microcrystalline cellulose, modified celluloses, hydroxypropyl cellulose, sodium carboxymethyl cellulose, alginic acid and polyvinyl pyrrolidone; 10. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one disintegrant selected from the group consisting of dibasic calcium phosphate, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, corn starch, alginic acid, hydroxypropylcellulose, calcium carboxymethylcellulose, sodium carboxymethylcellulose, cross-linked sodium carboxymethylcellulose, ion exchange swelling resins, alginates, formaldehyde-casein, cellulose, croscarmellose sodium, crospovidone, cellulose mi- crocrystalline, sodium carboxymethyl starch, sodium starch glycolate, starches and rice starch. 12. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one lubricant selected from the group consisting of calcium stearate, magnesium stearate, sodium stearyl fumarate, stearic acid, stearate zinc, talc and waxes. 13. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one non-stick is selected from the group consisting of colloidal silica dioxide and talc.; and 14. Composition according to claim 7, characterized in that the at least one pharmaceutically acceptable excipient is at least one diluent selected from the group consisting of mannitol, sucrose, anhydrous dibasic calcium phosphate, dibasic calcium phosphate dihydrate - anhydrous co, tribasic calcium phosphate, cellulose, lactose, magnesium carbonate and microcrystalline cellulose. 15. Dosage form, characterized in that it comprises the composition, as defined in claim 1, comprising at least one digestive enzyme as a plurality of coated digestive enzyme particles, wherein the at least one digestive enzyme is pancrelipase, wherein each said particle contains a core comprising pancrelipase coated with an enteric coating, and, that the enteric coating comprises an enteric polymer and further comprises 4 to 10% talc based on the total weight of the particle, wherein the composition has a moisture content of 3% or less. 16. Dosage form according to claim 15, characterized in that it is a tablet or capsule comprising the composition as defined in claim 1. 17. Dosage form according to claim 16, characterized in that said capsule is filled with the composition as defined in claim 1. 18. A dosage form according to claim 16, characterized in that it comprises: a capsule filled with a plurality of coated particles; the coated particles comprise an enteric coated core; the core comprises pancrelipase and at least one disintegrant; and the enteric coating comprises at least one polymer and 20 to 60% by weight of talc, based on the total weight of the coating; wherein the coated particles have a content of 3% or less, and the capsule has a moisture content of 4% or less. 19. Composition according to claim 1, characterized in that the enteric polymer is selected from the group consisting of cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, copolymers of methacrylic acid-methylmethacrylate and shellac. 20. Composition according to claim 1, characterized in that the enteric coating further comprises a plasticizer. 21. Composition according to claim 20, characterized in that the enteric coating comprises 10 to 20% by weight of at least one enteric polymer and 1 to 2% of at least one plasticizer based on the total weight of the particles coated. 22. Composition according to claim 20, characterized in that the plasticizer is selected from the group consisting of triacetin, tributyl citrate, triethyl citrate, acetyl tri-n-butyl citrate, diethyl phthalate, sebacate dibutyl, polyethylene glycol, polypropylene glycol, castor oil, acetylated mono-glyceride, acetylated di-glyceride and mixtures thereof. 23. Dosage form according to claim 17 or 18, characterized in that the capsule has a water content of 2% or less. 24. Dosage form according to claim 17 or 18, characterized in that the capsule comprises a material selected from the group consisting of a cellulosic polymer, hydroxypropylmethylcellulose, starch, polysaccharide, pullulan and gelatin. 25. Dosage form according to claim 24, characterized in that the capsule comprises hydroxypropyl methylcellulose. 26. Dosage form according to claim 24, characterized in that the capsule comprises a material selected from the group consisting of a cellulosic polymer, hydroxypropylmethylcellulose, starch, polysaccharide, pullulan and gelatin and has a moisture content of less than 4% or less than 2%. 27. Dosage form according to claim 22, characterized in that the capsule comprises hydroxypropylmethylcellulose and has a moisture content of less than 4% or less than 2%. 28. Packaging, characterized in that it comprises a sealed container comprising a moisture resistant material, a desiccant and at least one dosage form as defined in claim 15, wherein the desiccant and at least one dosage form are within of the sealed container. 29. Packaging according to claim 28, characterized in that the moisture resistant material is selected from the group consisting of metal, glass, plastic and metal-coated plastic. 30. Packaging according to claim 28, characterized in that the desiccant is selected from the group consisting of molecular sieve, clay, silica gel, activated carbon and combinations thereof. 31. Packaging according to claim 30, characterized in that the desiccant are molecular sieves. 32. Use of a composition as defined in claim 1, characterized in that it is for the preparation of a medicine to treat or prevent a disorder associated with digestive enzyme deficiency in a mammal in need thereof. 33. Method of preparing a composition according to claim 1, characterized in that it comprises: Coating particles of at least one digestive enzyme in an atmosphere having a moisture content of 3.6 g of water per m3 or less, with an enteric coating comprising an enteric polymer and talc, thus forming a plurality of delayed release dosage units. 34. Method according to claim 33, characterized in that the atmosphere comprises air, nitrogen or an inert gas. 35. Method according to claim 33, characterized in that the digestive enzyme particles are coated with a mixture of an enteric polymer dissolved in acetone and talc. 36. Composition according to claim 1, characterized in that the pancrelipase is derived from pig. 37. Dosage form, characterized in that it is a dosage form with zero excess filling comprising the composition as defined in claim 1. 38. Use of an effective amount of the composition as defined in claim 1, characterized in that in the preparation of a medicament for the treatment of exocrine pancreatic insufficiency in a patient. 39. Use according to claim 38, characterized in that the patient has cystic fibrosis. 40. Use according to claim 38, characterized in that said treatment alleviates the malabsorption of fat in a patient. 41. Use according to claim 38, characterized in that said treatment increases the fat absorption coefficient in a patient. 42. Use according to claim 41, characterized in that said treatment increases the fat absorption coefficient by at least 85% in a patient. 43. Use according to claim 38, characterized in that said increase in the coefficient of fat absorption in a patient occurs without co-administration of a proton pump inhibitor.